蝴蝶兰成花差异基因的筛选与分析

Screening and Analysis of Differential Genes in Flowering Stages of Phalaenopsis

为探明蝴蝶兰花芽分化调控的内在分子机制,以及花芽分化过程中的关键基因,本研究以蝴蝶兰不同分化时期的花芽进行转录组测序,筛选调控植株成花转变的关键基因并进行实时荧光定量PCR (qRT-PCR)验证。测序共得到44.42 Gb的原始数据,分别注释到7个公共数据库(Swiss-Prot, Nr, KEGG, KOG, Pfam,eggNOG和GO)。其中成功被GO注释到的Unigenes有16 181条;被KEGG注释到的Unigenes共有8 099条,分别注释到207个代谢通路。差异表达基因分析表明,2个花芽分化时期的蝴蝶兰共获得6 088条差异表达基因,其中上调表达有3 319条,下调表达有2 769条。通过查阅文献寻找出36个与蝴蝶兰花芽分化有关的基因。光周期调节途径中得到CRY1和PHYA;生物节律相关基因PIF4和EMF1;年龄途径相关基因SPL等;以及开花整合因子基因CO、FT、LFY等。选取GA序列、FD序列、FT序列、CO序列4个相关基因进行q RT-PCR验证,结果与测序结果相符。本研究为蝴蝶兰花芽调控提供一定的理论指导。

In order to clarify the internal molecular mechanism of phalaenopsis orchid bud differentiation regulation and key genes during flower bud differentiation.High-throughput sequencing technology was used to sequence the transcriptomes of different stages of flower bud differentiation to screen the key genes that regulate plant flower formation and perform validated by real-time quantitative PCR(qRT-PCR).A total of 44.42 Gb of raw data was obtained by sequencing and annotated to 7 public databases(Swiss-Prot,Nr,KEGG,KOG,Pfam,eggNOG,and GO).Among them,there were 16181 unigenes successfully annotated by GO;8099 unigenes annotated by KEGG were annotated to 207 metabolic pathways respectively.Analysis of differentially expressed genes showed that 6088 differentially expressed genes were obtained in the two flower bud differentiation stages,of which 3319 were up-regulated and 2769 were down-regulated.Through literature review,36 genes related to Phalaenopsis bud differentiation were identified.CRY1 and PHYA are obtained in the photocycle regulation pathway.Biorhythmrelated genes PIF4 and EMF1;age pathway-related genes SPL,etc.;and flowering integration factor genes:CO,FT,LFY,etc..GA-,FD-,FT-,and CO-related genes were selected for qRT-PCR and the results were consistent with the sequencing results.This study provided some theoretical guidance for the regulation of Phalaenopsis orchid buds.